Neuronal polarization occurs shortly after mitosis. In neurons differentiating in vitro, axon formation follows the segregation of growth-promoting activities to only one of the multiple neurites that form after mitosis1,2. It is unresolved whether such spatial restriction makes use of an intrinsic program, like during C. elegans embryo polarization3, or is extrinsic and cue-mediated, as in migratory cells4. Here we show that in hippocampal neurons in vitro, the axon consistently arises from the neurite that develops first after mitosis. Centrosomes, the Golgi apparatus and endosomes cluster together close to the area where the first neurite will form, which is in turn opposite from the plane of the last mitotic division. We show that the polarized activities of these organelles are necessary and sufficient for neuronal polarization: (1) polarized microtubule polymerization and membrane transport precedes first neurite formation, (2) neurons with more than one centrosome sprout more than one axon and (3) suppression of centrosome-mediated functions precludes polarization. We conclude that asymmetric centrosome-mediated dynamics in the early post-mitotic stage instruct neuronal polarity, implying that pre-mitotic mechanisms with a role in division orientation may in turn participate in this event.
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da Silva, J. S. & Dotti, C. G. Breaking the neuronal sphere: regulation of the actin cytoskeleton in neuritogenesis. Nature Rev. Neurosci. 3, 694–704 (2002)
Horton, A. C. & Ehlers, M. D. Neuronal polarity and trafficking. Neuron 9, 277–295 (2003)
Cowan, C. R. & Hyman, A. A. Centrosomes direct cell polarity independently of microtubule assembly in C. elegans embryos. Nature 431, 92–96 (2004)
Singer, S. J. & Kupfer, A. The directed migration of eukaryotic cells. Annu. Rev. Cell Biol. 2, 337–365 (1986)
Dotti, C. G., Sullivan, C. A. & Banker, G. The establishment of polarity by hippocampal neurons in culture. J. Neurosci. 8, 1454–1468 (1988)
Bradke, F. & Dotti, G. C. The role of actin instability in axon formation. Science 283, 1931–1934 (1999)
Ueda, M., Graf, R., MacWilliams, H. K., Schliwa, M. & Euteneuer, U. Centrosome positioning and directionality of cell movements. Proc. Natl Acad. Sci. USA 94, 9674–9678 (1997)
Grill, S. W., Howard, J., Schaffer, E., Stelzer, E. H. & Hyman, A. A. The distribution of active force generators controls mitotic spindle position. Science 301, 518–521 (2003)
Dotti, C. G. & Banker, G. Intracellular organization of hippocampal neurons during the development of neuronal polarity. J. Cell Sci. 15 (suppl.), 75–84 (1991)
Yonemura, E. M. et al. Rho-dependent transfer of citron-kinase to the cleavage furrow of dividing cells. J. Cell Sci. 114, 3273–3284 (2001)
Wu, C. F., Suzuki, N. & Poo, M. M. Dissociated neurons from normal and mutant Drosophila larval central nervous system in cell culture. J. Neurosci. 3, 1888–1899 (1983)
Megraw, T. L., Kilaru, S., Turner, F. R. & Kaufman, T. C. The centrosome is a dynamic structure that ejects PCM flares. J. Cell Sci. 115, 4707–4718 (2002)
Carney, G. E., Wade, A. A., Sapra, R., Goldstein, E. S. & Bender, M. DHR3, an ecdysone-inducible early-late gene encoding a Drosophila nuclear receptor, is required for embryogenesis. Proc. Natl Acad. Sci. USA 94, 12024–12029 (1997)
Houliston, E. & Maro, B. Post-translational modification of distinct microtubule subpopulations during cell polarization and differentiation in the mouse preimplantation embryo. J. Cell Biol. 108, 543–551 (1989)
Surrey, T. et al. Chromophore-assisted light inactivation and self-organization of microtubules and motors. Proc. Natl Acad. Sci. USA 14, 4293–4298 (1998)
Sydor, A. M., Su, A. L., Wang, F. S., Xu, A. & Jay, D. G. Talin and vinculin play distinct roles in filopodial motility in the neuronal growth cone. J. Cell Biol. 134, 1197–1207 (1996)
Wu, C. F., Sakai, K., Saito, M. & Hotta, Y. Giant Drosophila neurons differentiated from cytokinesis-arrested embryonic neuroblasts. J. Neurobiol. 21, 499–507 (1990)
Craig, A. M. & Banker, G. Neuronal polarity. Annu. Rev. Neurosci. 17, 267–310 (1994)
Yoshimura, T. et al. GSK-3β regulates phosphorylation of CRMP-2 and neuronal polarity. Cell 14, 137–149 (2005)
Shi, S. H., Cheng, T., Jan, L. Y. & Jan, Y. N. APC and GSK-3β are involved in mPar3 targeting to the nascent axon and establishment of neuronal polarity. Curr. Biol. 23, 2025–2032 (2004)
Schwamborn, J. C. & Puschel, A. W. The sequential activity of the GTPases Rap1B and Cdc42 determines neuronal polarity. Nature Neurosci. 7, 923–929 (2004)
Nishimura, T. et al. Role of the PAR-3–KIF3 complex in the establishment of neuronal polarity. Nature Cell Biol. 6, 328–334 (2004)
Shi, S. H., Jan, L. Y. & Jan, Y. N. Hippocampal neuronal polarity specified by spatially localized mPar3/mPar6 and PI 3-kinase activity. Cell 10, 63–75 (2003)
Goslin, K. & Banker, G. Experimental observations on the development of polarity by hippocampal neurons in culture. J. Cell Biol. 108, 1507–1516 (1989)
da Silva, J. S., Hasegawa, T., Miyagi, T., Dotti, C. G. & Abad-Rodríguez, J. Asymmetric membrane ganglioside sialidase activity specifies axonal fate. Nature Neurosci. 8, 606–615 (2005)
Piperno, G. & Fuller, M. T. Monoclonal antibodies specific for an acetylated form of α-tubulin recognize antigens in cilia and flagella from a variety of organisms. J. Cell Biol. 101, 2085–2094 (1985)
De Hoop, M., Meyn, L. & Dotti, C. G. in Methods. in Cell Biology: Laboratory Handbook 2nd edn (ed. Celis, J. E.) 154–163 (Academic Press, San Diego, 1998)
Feiguin, F., Llamazares, S. & Gonzalez, C. Methods in Drosophila cell cycle biology. Curr. Top. Dev. Biol. 36, 279–291 (1998)
We would like to thank E. Cassin and B. Hellias for the hippocampal neurons, L. Ciapponi and the Bloomington Stock Center for fly strains, and C. Gonzalez for advice with the neuroblast division experiment. F.C.dA. is supported by an EMBO long-term fellowship. J.S.D.S. was supported by an FCT/PRAXIS XXI scholarship (Portuguese Ministry of Science and Technology). F.F. was supported by an Alexander von Humboldt scholarship. Part of this work is supported by an EU Contract grant (APOPIS) to C.G.D.Author Contributions F.C.d.A. and G.P. were responsible for all in vitro experiments in mammalian and insect neurons, respectively. J.S.D.S. supervised the hippocampal neuron work. P.G.C. helped with the in situ work.
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
The first sprout contains neuronal polarity information. (PDF 229 kb)
Organelle polarization marks the site of neuronal polarity. (PDF 496 kb)
Drosophila neurons' polarization in vitro and in situ correlates with plane of mitotic division and the localization of the centrosomes. (PDF 592 kb)
Centrosomal-mediated polarized microtubule and membrane activities precede morphological polarization in vitro and in situ. (PDF 177 kb)
Pharmacological disruption of microtubule polymerization and membrane trafficking prevents morphological polarization. (PDF 99 kb)
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de Anda, F., Pollarolo, G., Da Silva, J. et al. Centrosome localization determines neuronal polarity. Nature 436, 704–708 (2005). https://doi.org/10.1038/nature03811
Neuroscience Bulletin (2022)
Molecular Neurodegeneration (2018)
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